Neuropeptide Y Expression Levels Research Articles

FunctionalNPYVariation as a Factor in Stress Resilience and Alcohol Consumption in Rhesus Macaques

Neuropeptide Y (NPY) counters stress and is involved in neuroadaptations that drive escalated alcohol drinking in rodents. In humans, low NPY expression predicts amygdala response and emotional reactivity. Genetic variation that affects the NPY system could moderate stress resilience and susceptibility to alcohol dependence. To determine whether functional NPY variation influences behavioral adaptation to stress and alcohol consumption in a nonhuman primate model of early adversity (peer rearing). We sequenced the rhesus macaque NPY locus (rhNPY) and performed in silico analysis to identify functional variants. We performed gel shift assays using nuclear extract from testes, brain, and hypothalamus. Levels of NPY in cerebrospinal fluid were measured by radioimmunoassay, and messenger RNA levels were assessed in the amygdala using real-time polymerase chain reaction. Animals were exposed to repeated social separation stress and tested for individual differences in alcohol consumption. Animals were genotyped for -1002 T > G, and the data were analyzed using analysis of variance. National Institutes of Health Animal Center. Subjects Ninety-six rhesus macaques. Main Outcome Measure Behavior arousal during social separation stress and ethanol consumption. The G allele altered binding of regulatory proteins in all nuclear extracts tested, and -1002 T > G resulted in lower levels of NPY expression in the amygdala. Macaques exposed to adversity had lower cerebrospinal fluid NPY levels and exhibited higher levels of arousal during stress, but only as a function of the G allele. We also found that stress-exposed G allele carriers consumed more alcohol and exhibited an escalation in intake over cycles of alcohol availability and deprivation. Our results suggest a role for NPY promoter variation in the susceptibility to alcohol use disorders and point to NPY as a candidate for examining gene x environment interactions in humans.

Neuropeptide Y Knockout Mice Reveal a Central Role of NPY in the Coordination of Bone Mass to Body Weight

Changes in whole body energy levels are closely linked to alterations in body weight and bone mass. Here, we show that hypothalamic signals contribute to the regulation of bone mass in a manner consistent with the central perception of energy status. Mice lacking neuropeptide Y (NPY), a well-known orexigenic factor whose hypothalamic expression is increased in fasting, have significantly increased bone mass in association with enhanced osteoblast activity and elevated expression of bone osteogenic transcription factors, Runx2 and Osterix. In contrast, wild type and NPY knockout (NPY −/−) mice in which NPY is specifically over expressed in the hypothalamus (AAV-NPY+) show a significant reduction in bone mass despite developing an obese phenotype. The AAV-NPY+ induced loss of bone mass is consistent with models known to mimic the central effects of fasting, which also show increased hypothalamic NPY levels. Thus these data indicate that, in addition to well characterized responses to body mass, skeletal tissue also responds to the perception of nutritional status by the hypothalamus independently of body weight. In addition, the reduction in bone mass by AAV NPY+ administration does not completely correct the high bone mass phenotype of NPY −/− mice, indicating the possibility that peripheral NPY may also be an important regulator of bone mass. Indeed, we demonstrate the expression of NPY specifically in osteoblasts. In conclusion, these data identifies NPY as a critical integrator of bone homeostatic signals; increasing bone mass during times of obesity when hypothalamic NPY expression levels are low and reducing bone formation to conserve energy under ‘starving’ conditions, when hypothalamic NPY expression levels are high.

Cloning, distribution and effects of season and nutritional status on the expression of neuropeptide Y (NPY), cocaine and amphetamine regulated transcript (CART) and cholecystokinin (CCK) in winter flounder (Pseudopleuronectes americanus)

cDNAs encoding for neuropeptide Y (NPY), cocaine and amphetamine regulated transcript (CART) and cholecystokinin (CCK) were cloned in winter flounder, a species that undergoes a period of natural fasting during the winter. Tissue distribution studies show that these peptides are present in several peripheral tissues, including gut and gonads, as well as within the brain. We assessed the effects of season and fasting on the expression of these peptides. Our results show that NPY and CCK, but not CART, show seasonal differences in expression with higher hypothalamic NPY and lower gut CCK expression levels in the winter. In the summer, fasting induced an increase in hypothalamic NPY expression levels and a decrease in gut CCK levels, but did not affect hypothalamic CART expression levels. None of the peptides examined was affected by fasting in the winter. Our results suggest that NPY and CCK, but maybe not CART, might have a major role in the regulation of feeding in winter flounder and might contribute to the seasonal fluctuations in appetite in this species.

Difference of NPY and its Receptor Gene Expressions between Obesity and Obesity-resistant Rats in Response to High-fat Diet

To investigate the effect of high-fat diet on expressions of hypothalamic neuropeptide Y (NPY) and Y1, Y2, Y5 receptors and the mechanism relative to the susceptibility of obesity. Thirty-six female SD rats were randomly divided into high-fat diet group (HF, n=27) and chow food group (CF, n=9), and given either HF or CF diet for 13 weeks. Then the HF diet group was subdivided into dietary induced obesi-ty (DIO) and dietary induced obesity resistant (DIO-R) rat according to the final body weight. Body weight, caloric intake, energy efficiency, visceral fat pads were measured and compared. The levels of plasma and hypothalamic NPY were determined by Radioimmunoassay. Real-time Polymerase Chain Reaction (PCR) was used to measure the gene expression of NPY and its receptors in hypothalamus. Body weight, caloric intake, energy efficiency and visceral fat pads in DIO rats were higher significantly than those in CF or DIO-R rats (p<0.01). No difference in plasma NPY level was detected among the three groups (p>0.05); the hypothalamic NPY level was significantly higher in DIO group than in CF or DIO-R group (p<0.01). Gene expression levels of NPY and Y1, Y2, Y5 receptors were higher significantly in DIO group than in CF or DIO-R group (p<0.01), while no significant difference was found between DIO-R and CF rats (p>0.05), except that Y2 receptor was lower in DIO-R rats (p<0.01). SD rat fed with a HF diet showed different susceptibility to obesity, and up-regulated hypothalamic NPY and Y1, Y2, Y5 receptor gene expressions were closely associated with being predisposed to obesity and overeating of DIO rats.

High fat diet induced hepatic insulin resistance is not related to changes in hypothalamic mRNA expression of NPY, AgRP, POMC and CART in mice

The hypothalamic circuitry, apart from its impact on food intake, modulates insulin sensitivity to adapt metabolic conditions in the face of environmental fluctuations in nutrient availability. The purpose of the present study was to investigate the effects of 2 weeks high fat feeding in wildtype mice on (1) insulin sensitivity and triglyceride accumulation in liver and muscle in relation to (2) mRNA expression levels of Neuropeptide Y (NPY), Agouti-related protein (AgRP), pro-opiomelanocortin (POMC), and cocaine- and amphetamine-regulated transcript (CART) in the hypothalamus. Two weeks of high fat feeding induced hepatic insulin resistance in the presence of increased hepatic triglyceride accumulation. In muscle, however, 2 weeks of high fat feeding did not result in changes in insulin sensitivity or in triglyceride content. mRNA expression levels of NPY, AgRP, POMC, and CART in the hypothalamus were not different between the groups. This study shows that 2 weeks of high fat feeding in mice does not affect mRNA expression levels of NPY, AgRP, POMC or CART, in the whole hypothalamus, despite induction of hepatic, but not peripheral, insulin resistance. Therefore, a major physiological role of these neuroendocrine factors in the induction of hepatic insulin resistance during a high fat diet seems less likely.

Expression of neuropeptide Y by glutamatergic stimulation in rat C6 glioma cells

We have investigated the expression of neuropeptide Y (NPY) in C6 glioma cells after the glutamatergic stimulation by the in situ RT-PCR and immunocytochemical techniques. The expression of NPY mRNA correlated well with immunocytological findings in each series of experiments. NPY protein expression was enhanced by glutamate (1, 10, 50, 100 μM, and 1 mM) dose-dependently, and its expression was slightly increased by N-methyl- d-aspartate (NMDA; 1, 10, 100, 500 μM, and 1 mM) and kainic acid (1, 10, 100, 300 μM, and 1 mM). We pretreated the cells with dopamine, haloperidol, pentylenetetrazol, and muscimol before each stimulation. The pentylenetetrazol and muscimol did not significantly alter the patterns of NPY expression induced by the glutamatergic stimulation. On the other hand, the dopamine and haloperidol pretreatment significantly elevated the levels of NPY expression that were induced by NMDA and kainic acid. Our results indicate that NPY release is closely related to glutamatergic stimulation, and it could be dynamically mediated by GABAergic and dopaminergic costimulation.

Alterations in neuropeptide Y and Y1 receptor mRNA expression in brains from an animal model of depression: region specific adaptation after fluoxetine treatment

To investigate the possible link between neuropeptide Y (NPY) and depression, we analyzed NPY and its receptors in different limbic-related regions in the Flinder sensitive line (FSL), a genetic animal model of depression. In situ hybridization histochemistry was used to measure mRNA expression levels of NPY and NPY receptors, Y1 and Y2, in the FSL as compared to the control Flinder resistant Line rats (FRL). In the FSL rats, NPY mRNA expression levels were significantly decreased in the nucleus accumbens and CA regions, but increased in the arcuate nucleus and anterior cingulate cortex. Y1 receptor mRNA expression was decreased in different cortical regions (retrosplenial, anterior cingulate, and occipital) and in the hippocampal dentate gyrus. Y2 mRNA expression levels did not differ between FSL and FRL animals. The effect of the antidepressant drug fluoxetine (a serotonin reuptake inhibitor) in the two rat strains was also studied. There was an increase of the NPY mRNA hybridization signal in the arcuate nucleus of both strains following the antidepressant treatment (10 μmol/kg; daily for 14 days). However, in other brain regions, fluoxetine administration caused a differential effect on the induction of NPY-related genes in the two rat strains: in the CA region and dentate gyrus NPY mRNA expression was increased in the FSL, but decreased in the FRL. In contrast, Y1 mRNA levels tended to be decreased by fluoxetine in the nucleus accumbens of the FSL rats, but increased in the FRL. These findings suggest an involvement of the Y1, but not the Y2, receptor subtype in depressive disorder. Overall, the results appear to sustain the importance of the FSL rats as an animal model of depression in view of the impairment of NPY genes and the ability of fluoxetine treatment to normalize NPY-related gene expression selectively in this strain.

Temporal analysis of neuropeptide Y expression in the rat trigeminal ganglion following peripheral axotomy of the inferior alveolar nerve

Temporal changes in the appearance and the cell-size spectrum of neuropeptide Y (NPY)-like immunoreactive (IR) cells in the rat trigeminal ganglion (TG) following peripheral axotomy of the inferior alveolar nerve (IAN) were studied by retrograde neuronal tracing with True Blue (TB) and immunohistochemistry for NPY. The number of labeled cells following application of TB to the cut-end of the IAN increased rapidly up to 3 days, and was maintained at a constant level thereafter. The size distribution of cross-sectional areas of TB-labeled cells was similar at 3 days and afterwards. NPY-IR cells, which were not detected in the normal TG, appeared on the first day following axotomy, and increased gradually in number reaching a maximum at 14 days. The frequency histogram of the cross-sectional areas of NPY-IR cells was similar at 3 days and afterwards. The present results indicate that the effect of nerve injury on the levels of NPY expression in the sensory neurons began soon after nerve injury, reaching a maximum around 14 days, and that induction of NPY in the sensory neurons occurred in the same cell size-specific manner for a long period.

Food deprivation and age modulate neuropeptide gene expression in the murine hypothalamus and adrenal gland

We have developed a rapid and simple method for quantitative Northern blot analysis of rare messenger RNA species from single mouse hypothalami and adrenals. This technique has allowed us to study the effects of food deprivation on neuropeptide gene expression in the mouse hypothalamus and adrenal gland. The potential modulatory effects of sex and age were also investigated. Food deprivation induced a two-fold increase in the amount of hypothalamic neuropeptide Y (NPY) mRNA, but did not increase NPY mRNA in the brainstem. Age had a significant effect on levels of NPY messenger RNA levels in the hypothalamus. However, there were no gender-associated effects. Sexually immature females (6 weeks old) had higher levels of NPY expression than mature females (9 weeks old). In contrast, in the adrenal gland, increasing maturity was associated with higher levels of NPY mRNA. As in the hypothalamus, fasting caused approximately two-fold increases in NPY mRNA over levels in the ad libitum fed state for both mature and immature mice. Thus, hypothalamic NPY neurons are responsive to nutritional deprivation and developmental status, suggesting that NPY neurons may be important in energy homeostasis.